The vertebrate ??crystallins play an essential role in lens development and the maintenance of lens transparency. Abnormal expression of ??crystallins is also involved in numerous human diseases of the eye, muscular and nervous systems, and has recently been implicated in various cancers. Our goals in this proposal are to leverage the benefits of the zebrafish as a model system to 1) detail the roles of ??crystallins in lens development and 2) identify novel regulatory regions in the mouse and zebrafish 1?? -crystallin promoter. These studies will contribute to our understanding of normal and pathological lens function. We propose testing the hypothesis that the role of ??crystallins in lens development is conserved between zebrafish and mammals by using synthetic anti-sense oligonucleotide molecules called morpholinos to inhibit translation of each zebrafish ??crystallin individually and in combination. This work will provide an efficient model system for analyzing the specifics of ?? crystallin developmental function, and will allow future in vivo studies of genetically modified ??crystallins. The unusual presence of two 1?? -crystallins in zebrafish (one lens specific and one ubiquitously expressed in the body) will allow us to examine how the functions of mammalian 1?-crystallin have become divided between the two zebrafish paralogs. The zebrafish will also be used as a rapid screening system to identify novel regulatory regions controlling 1?? -crystallin's temporal and spatial expression. Regulatory elements are so far only known from a relatively small proximal region of the mammalian 1?? -crystallin promoter. Future studies can utilize this zebrafish model to test the in vivo effects of mutated promoters on ??crystallin expression. The proposed work will be done in an undergraduate biology department, allowing students to gain experience in experimental design, data analysis and cutting edge research techniques used in visual science. PUBLIC HEALTH RELEVANCE: ??crystallin proteins are necessary for eye lens development and the maintenance of lens transparency. Mutations in these proteins and abnormal amounts are associated with multiple diseases of the eye and nervous system and found in various cancers. This project will use the zebrafish as a model species for examining the role of ??crystallins in lens development and the mechanisms that control their production.